Coherent diffractive imaging of microtubules using an X-ray laser

Nat Commun. 2019 Jun 13;10(1):2589. doi: 10.1038/s41467-019-10448-x.

Abstract

X-ray free electron lasers (XFELs) create new possibilities for structural studies of biological objects that extend beyond what is possible with synchrotron radiation. Serial femtosecond crystallography has allowed high-resolution structures to be determined from micro-meter sized crystals, whereas single particle coherent X-ray imaging requires development to extend the resolution beyond a few tens of nanometers. Here we describe an intermediate approach: the XFEL imaging of biological assemblies with helical symmetry. We collected X-ray scattering images from samples of microtubules injected across an XFEL beam using a liquid microjet, sorted these images into class averages, merged these data into a diffraction pattern extending to 2 nm resolution, and reconstructed these data into a projection image of the microtubule. Details such as the 4 nm tubulin monomer became visible in this reconstruction. These results illustrate the potential of single-molecule X-ray imaging of biological assembles with helical symmetry at room temperature.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Algorithms
  • Crystallography, X-Ray / instrumentation
  • Crystallography, X-Ray / methods
  • Electrons*
  • Image Processing, Computer-Assisted
  • Lasers*
  • Microtubules / ultrastructure*
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods*
  • Scattering, Radiation
  • Synchrotrons
  • Tubulin / ultrastructure*
  • X-Rays

Substances

  • Tubulin